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dc.contributor.authorKiehl, Mariana Fitarellipt_BR
dc.contributor.authorMacedo, Gabriel de Souzapt_BR
dc.contributor.authorSchlatter, Rosane Paixãopt_BR
dc.contributor.authorSantos, Patrícia Koehler dospt_BR
dc.contributor.authorMatte, Ursula da Silveirapt_BR
dc.contributor.authorProlla, Patrícia Ashtonpt_BR
dc.contributor.authorGiacomazzi, Julianapt_BR
dc.date.accessioned2016-08-20T02:14:58Zpt_BR
dc.date.issued2016pt_BR
dc.identifier.issn1415-4757pt_BR
dc.identifier.urihttp://hdl.handle.net/10183/147480pt_BR
dc.description.abstractGermline mutations in the TP53 gene are associated with Li-Fraumeni and Li-Fraumeni-Like Syndromes, characterized by increased predisposition to early-onset cancers. In Brazil, the prevalence of the TP53-p.R337H germline mutation is exceedingly high in the general population and in cancer-affected patients, probably as result of a founder effect. Several genotyping methods are used for the molecular diagnosis of LFS/LFL, however Sanger sequencing is still considered the gold standard. We compared performance, cost and turnaround time of Sanger sequencing, PCR-RFLP, TaqMan-PCR and HRM in the p.R337H genotyping. The performance was determined by analysis of 95 genomic DNA samples and results were 100% concordant for all methods. Sequencing was the most expensive method followed by TaqMan-PCR, PCR-RFLP and HRM. The overall cost of HRM increased with the prevalence of positive samples, since confirmatory sequencing must be performed when a sample shows an abnormal melting profile, but remained lower than all other methods when the mutation prevalence was less than 2.5%. Sequencing had the highest throughput and the longest turnaround time, while TaqMan-PCR showed the lowest turnaround and hands-on times. All methodologies studied are suitable for the detection of p.R337H and the choice will depend on the application and clinical scenario.en
dc.format.mimetypeapplication/pdfpt_BR
dc.language.isoengpt_BR
dc.relation.ispartofGenetics and molecular biology. Ribeirão Preto, SP. Vol. 39, no. 2 (Jun. 2016), p. 203-209pt_BR
dc.rightsOpen Accessen
dc.subjectTP53-p.R337Hen
dc.subjectNeoplasiaspt_BR
dc.subjectPredisposição genética para doençapt_BR
dc.subjectRFLPen
dc.subjectTaqManen
dc.subjectHRMen
dc.subjectSanger sequencingen
dc.titleComparison of multiple genotyping methods for the identification of the cancer predisposing founder mutation p.R337H in TP53pt_BR
dc.typeArtigo de periódicopt_BR
dc.identifier.nrb000997097pt_BR
dc.type.originNacionalpt_BR


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